Analysis
A variety of methods used in detection, purification, and structural analysis of glycoproteins are
| Method | Use |
|---|---|
| Periodic acid-Schiff stain | Detects glycoproteins as pink bands after electrophoretic separation. |
| Incubation of cultured cells with glycoproteins as radioactive decay bands | Leads to detection of a radioactive sugar after electrophoretic separation. |
| Treatment with appropriate endo- or exoglycosidase or phospholipases | Resultant shifts in electrophoretic migration help distinguish among proteins with N-glycan, O-glycan, or GPI linkages and also between high mannose and complex N-glycans. |
| Agarose-lectin column chromatography, lectin affinity chromatography | To purify glycoproteins or glycopeptides that bind the particular lectin used. |
| Lectin affinity electrophoresis | Resultant shifts in electrophoretic migration help distinguish and characterize glycoforms, i.e. variants of a glycoprotein differing in carbohydrate. |
| Compositional analysis following acid hydrolysis | Identifies sugars that the glycoprotein contains and their stoichiometry. |
| Mass spectrometry | Provides information on molecular mass, composition, sequence, and sometimes branching of a glycan chain. |
| NMR spectroscopy | To identify specific sugars, their sequence, linkages, and the anomeric nature of glycosidic chain. |
| Multi-angle light scattering | In conjunction with size-exclusion chromatography, UV/Vis absorption and differential refractometry, provides information on molecular mass, protein-carbohydrate ratio, aggregation state, size, and sometimes branching of a glycan chain. In conjunction with composition-gradient analysis, analyzes self- and hetero-association to determine binding affinity and stoichiometry with proteins or carbohydrates in solution without labeling. |
| Dual Polarisation Interferometry | Measures the mechanisms underlying the biomolecular interactions, including reaction rates, affinities and associated conformational changes. |
| Methylation (linkage) analysis | To determine linkage between sugars. |
| Amino acid or cDNA sequencing | Determination of amino acid sequence. |
Read more about this topic: Glycoprotein
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